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pLVX-TetOne™-Puro

All-in-one lentiviral vector with a puromycin resistance marker, for strong inducible expression of genes using the Tet-On® system.

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pLVX-TetOne-Puro Sequence and MappLVX-TetOne-Puro.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SgrDI (8859) SspI (8742) PvuI (8308) DrdI (7156) PciI (7048) BmtI (6664) NheI (6660) KpnI (5878) Acc65I (5874) Bsu36I (5870) PstI (5754) PflMI (5390) MluI (5136) BstEII (4670) RsrII (4652) FseI (1150) BbvCI - Bpu10I (1424) AleI (1577) KflI - PpuMI (1934) cPPT/CTS BamHI (2496) AgeI - SgrAI (2508) BstZ17I (2516) EcoRI (2522) PaeR7I - PspXI - XhoI (2893) BstXI (2915) SpeI (3168) AscI (3257) XbaI (3444) BfuAI - BspMI (3998) NdeI (4020) SfiI (4465) AvrII (4512) PflFI - Tth111I (4578) BsiWI (4592) pLVX-TetOne™-Puro 9227 bp
SgrDI  (8859)
1 site
C G T C G A C G G C A G C T G C
SspI  (8742)
1 site
A A T A T T T T A T A A
PvuI  (8308)
1 site
C G A T C G G C T A G C
DrdI  (7156)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PciI  (7048)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BmtI  (6664)
1 site
G C T A G C C G A T C G
NheI  (6660)
1 site
G C T A G C C G A T C G
KpnI  (5878)
1 site
G G T A C C C C A T G G
Acc65I  (5874)
1 site
G G T A C C C C A T G G
Bsu36I  (5870)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
PstI  (5754)
1 site
C T G C A G G A C G T C
PflMI  (5390)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
MluI  (5136)
1 site
A C G C G T T G C G C A
BstEII  (4670)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
RsrII  (4652)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
FseI  (1150)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
BbvCI  (1424)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1424)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
AleI  (1577)
1 site
C A C N N N N G T G G T G N N N N C A C
KflI  (1934)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (1934)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
BamHI  (2496)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
AgeI  (2508)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
SgrAI  (2508)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
BstZ17I  (2516)
1 site
G T A T A C C A T A T G
EcoRI  (2522)
1 site
G A A T T C C T T A A G
PaeR7I  (2893)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2893)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2893)
1 site
C T C G A G G A G C T C
BstXI  (2915)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
SpeI  (3168)
1 site
A C T A G T T G A T C A
AscI  (3257)
1 site
G G C G C G C C C C G C G C G G
XbaI  (3444)
1 site
T C T A G A A G A T C T
BfuAI  (3998)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (3998)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
NdeI  (4020)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
SfiI  (4465)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AvrII  (4512)
1 site
C C T A G G G G A T C C
PflFI  (4578)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (4578)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BsiWI  (4592)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
AmpR
7865 .. 8725  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7865 .. 8656  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7865 .. 8725  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   8657 .. 8725  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7865 .. 8725  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
Tet-On® 3G
3441 .. 4187  =  747 bp
248 amino acids  =  27.7 kDa
Product: modified rtTA protein that binds tightly to
promoters containing the tet operator in the
presence of doxycycline
Tet-On® 3G
3441 .. 4187  =  747 bp
248 amino acids  =  27.7 kDa
Product: modified rtTA protein that binds tightly to
promoters containing the tet operator in the
presence of doxycycline
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5945 .. 6578  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5945 .. 6578  =  634 bp
3' long terminal repeat (LTR) from HIV-1
PuroR
4536 .. 5135  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
4536 .. 5135  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
5149 .. 5737  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
5149 .. 5737  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
7109 .. 7694  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7109 .. 7694  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hPGK promoter
2912 .. 3422  =  511 bp
human phosphoglycerate kinase 1 promoter
hPGK promoter
2912 .. 3422  =  511 bp
human phosphoglycerate kinase 1 promoter
TRE3GS promoter
2528 .. 2895  =  368 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On® 3G, modified to
eliminate binding sites for endogenous mammalian
transcription factors
TRE3GS promoter
2528 .. 2895  =  368 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On® 3G, modified to
eliminate binding sites for endogenous mammalian
transcription factors
SV40 promoter
4198 .. 4527  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4198 .. 4527  =  330 bp
SV40 enhancer and early promoter
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
SV40 poly(A) signal
2187 .. 2321  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2187 .. 2321  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8878 .. 9012  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8878 .. 9012  =  135 bp
SV40 polyadenylation signal
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus
type 1
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)
AmpR promoter
8726 .. 8830  =  105 bp
AmpR promoter
8726 .. 8830  =  105 bp
MCS
2496 .. 2527  =  32 bp
multiple cloning site
MCS
2496 .. 2527  =  32 bp
multiple cloning site
SV40 ori
4378 .. 4513  =  136 bp
SV40 origin of replication
SV40 ori
4378 .. 4513  =  136 bp
SV40 origin of replication
tet operator
2653 .. 2671  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2653 .. 2671  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2689 .. 2707  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2689 .. 2707  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2725 .. 2743  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2725 .. 2743  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2761 .. 2779  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2761 .. 2779  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2797 .. 2815  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2797 .. 2815  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2833 .. 2851  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2833 .. 2851  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2869 .. 2887  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2869 .. 2887  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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